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The formation mechanism of kidney stones is complex. It is generally recognized that abnormal urine conditions or renal tubular epithelial cell damage, together with other factors cause the formation of renal papillary subepithelial calcium plaques (Randall’s plaques) or stone crystals that block the renal tubules (Randall’s plugs), and then oversaturated crystals gathering on Randall's plaque or plug and forming stones. However, there are many pathophysiological changes and manifestations, such as renal papillary anchoring stones, renal papillary crypts, renal papillary tip erosion, and exogenous renal papilla Renal papillary lesions, which may be an early manifestations before the formation of kidney stones. The study of renal papillary calcium plaque is very important for the pathogenesis of kidney stones, as well as the prevention and treatment of patients with stones. By focusing on the development process of Randall plaque theory, the formation and transformation mechanism of Randall plaque, as well as the manifestations and clinical treatment of the above mentioned different types of renal papillary calcium plaque lesions, this article reviewed three aspects of stone formation, including Randall’s plaque, renal papillary lesions with stones, and renal papillary lesions related to stone.
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Objective:To study the effect of Pterostilbene on endoplasmic reticulum stress and apoptosis in human renal tubular epithelial cells (HK-2 cells) induced by oxalate.Methods:From January 2019 to January 2020, HK-2 cells were divided into a control group (cultured with normal medium), an oxalate group (cultured with a medium containing 4 mmol/L of oxalate), and an intervention group of Pterostilbene (containing 4 mmol/L of oxalate + Pterostilbene 5, 10, and 20 μmol/L mixed medium were cultured at the same time), and the following tests were performed after 12 hours of treatment. Pterostilbene (5, 10, and 20 μmol/L) intervention group for cell viability test, lactate dehydrogenase cytotoxicity test, reduced glutathione, superoxide dismutase, malondialdehyde, hydrogen peroxide enzyme, total antioxidant capacity detection experiments to explore the degree of oxidative damage, and Western blotting experiments to explore the protein expression of ATF6, GRP78, DDIT3, caspase12, Clevead caspase 3/9; Pterostilbene (10 μmol/L) intervention group to detect mitochondrial membrane potential, caspase 3 enzyme activity, apoptosis rate, reactive oxygen detection to detect the apoptosis, reactive oxygen level, and qRT-PCR to detect ATF6, GRP78, DDIT3 of cells mRNA expression.Results:CCK-8 and lactate dehydrogenase toxicity test results showed that the cell activity of the oxalate group was significantly lower than that of the control group [(45.6±3.1)% vs. 100.0%, P<0.001]; the lactate dehydrogenase [(330.2±11.1)U/L vs. (2.6±6.7) U/L, P<0.001] of the oxalate group was higher than that of the control group increased obviously; the cell viability[ (57.2±1.7)%, (67.2±3.4)%, (78.9±1.8)%] of Pterostilbene intervention group (5, 10, 20 μmol/L) significantly increased compared with oxalate group ( P<0.05); lactate dehydrogenase [(288.1±4.3)U/L, (260.9±5.5)U, (202.7±10.2)U/L] in Pterostilbene intervention group (5, 10, 20 μmol/L ) was significantly lower than oxalate group ( P<0.05). The results of the five biochemical indexes of malondialdehyde, reduced glutathione, total superoxide dismutase, catalase, and total antioxidant capacity showed that the cell damage state was consistent with the experimental results of CCK-8 and lactate dehydrogenase. The active oxygen test results showed that the oxalate group had a significantly higher active oxygen level (76.3±4.9 vs. 6.2±1.7, P<0.01); the active oxygen level (39.5±5.4) of the Pterostilbene intervention group(10 μmol/L) was significantly lower than oxalate group ( P<0.01). The flow cytometry and caspase3 enzyme activity showed an increase in apoptosis rate and caspase3 activity in line with the trend of reactive oxygen levels. Mitochondrial membrane potential results showed that the oxalate group had a significantly lower mitochondrial membrane potential (0.76±0.15 vs. 7.84±0.26, P<0.01), and the mitochondrial membrane potential (2.26±0.27) of the Pterostilbene intervention group (10 μmol/L) was significantly higher than oxalate group( P<0.01). Western blot analysis showed that the relative expression of ATF6, DDIT3, GRP78, caspase12 and Cleaved caspase3/9 protein in the oxalate group was significantly higher than that in the control group. The relative expression of ATF6, DDIT3, GRP78, caspase12, Cleaved caspase3/9 protein in the Pterostilbene intervention group was significantly lower than that in the oxalate group ( P<0.05). qRT-PCR results showed that the mRNA expression trends of ATF6, DDIT3 and GRP78 in the three groups were consistent with the results of Western blotting. Conclusion:Pterostilbene can effectively inhibit the endoplasmic reticulum stress and apoptosis of HK-2 cells induced by oxalate.
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Objective To explore the implementing process and application effect of risk management in blood transfusion compatibility testing.Methods 16 957 patients receiving transfusion therapy along with blood transfusion compatibility testing at our hospital between July,2013 and June,2015 were selected as the control group,without any risk control in place.19 011 patients receiving such therapy yet with blood transfusion compatibility testing between July, 2015 and June, 2017 were selected as the observation group,and managed by the risk management procedure.The risk incidence and satisfactory rate of doctors,nurses and patients were analyzed between the two groups.Results The risk incidence was zero in the observation group, and 0.09% in the control group, indicating the risk incidence rate in the observation group significantly lower than the control group(P<0.05).The satisfactory rate of doctors, nurses and patients in the observation group(98.33%)was significantly higher than the control group (71.25%)(P <0.05).Conclusions Implementing risk management procedure in blood transfusion compatibility testing may effectively prevent and reduce the risk incidence, enhance the satisfactory rate of doctors,nurses and patients,and ensure the clinical transfusion safety.
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Objective To investigate the method for isolating and identification exosomes from the HK-2 cells exposed to high oxalate.Methods HK-2 cells were cultured to serum-free culture medium and treated with oxalate at the concentration from 0 mmol/L to 10.00 mmol/L for 48 h.CCK-8 assays were performed to measure the cells proliferation.Combined the cell morphology,observed under inverted microscope with statistical analysis,we finally 2.00 mmol/L oxalic acid as the experimental concentration.The HK-2 cell was exposed to 2.00 mmol/L oxalate for 48 h.The supernatants were collected.Exosomes were isolated and purified from the supernatants by ultracentrifugation.Transmission electron microscopy (TEM) was used to observe the morphology of isolated exosomes.Particle size of exosomes were detected with Nanosight technology.Western blot analysis was used to examine the experession of HSP70,CD63.Results CCK-8 assay showed that the cell viability in each group,including (100.0 ± 4.0) ％ in 0 mmol/L group;(97.7 ± 1.5)％ in the 0.25 mmol/L group;(97.3 ±2.1)％ in the 0.50 mmol/L group;(87.7 ± 2.1) ％ in the 1.00 mmol/L group;(76.0 ± 1.0) ％ in the 2.00 mmol/L group;(58.1 ± 2.6) ％ in the 4.00 mmol/L group;(52.7 ± 1.5) ％ in the 5.00 mmol/L group;(37.7 ± 3.2) ％ in the 8.00 mmol/L group;(31.3 ±2.0)％ in the 10.00 mmol/L group.The isolated exosomes demonstrated round or oval shape under TEM.The peak particle size was 56 nm,which the overall mean particle size was 87 nm.Those and particles with a diameter between 30-150 nm accounted for 91.2％.In this experiment,The expression of HSP70,CD63 could be detected in the isolated exosomes.However,only the expression of HSP70 could be detected in the HK-2 cells.Conclusions Under the treatment of 2 mmol/l oxalate for 48 hours,Ultracentrifugation can be used to isolate and purify exosomes efficiently from the HK-2 cells.This is helpful for further study of exosome as mediator of cell-to-cell communication.
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Objective To investigate the method for isolating and identification exosomes from the HK-2 cells exposed to high oxalate.Methods HK-2 cells were cultured to serum-free culture medium and treated with oxalate at the concentration from 0 mmol/L to 10.00 mmol/L for 48 h.CCK-8 assays were performed to measure the cells proliferation.Combined the cell morphology,observed under inverted microscope with statistical analysis,we finally 2.00 mmol/L oxalic acid as the experimental concentration.The HK-2 cell was exposed to 2.00 mmol/L oxalate for 48 h.The supernatants were collected.Exosomes were isolated and purified from the supernatants by ultracentrifugation.Transmission electron microscopy (TEM) was used to observe the morphology of isolated exosomes.Particle size of exosomes were detected with Nanosight technology.Western blot analysis was used to examine the experession of HSP70,CD63.Results CCK-8 assay showed that the cell viability in each group,including (100.0 ± 4.0) ％ in 0 mmol/L group;(97.7 ± 1.5)％ in the 0.25 mmol/L group;(97.3 ±2.1)％ in the 0.50 mmol/L group;(87.7 ± 2.1) ％ in the 1.00 mmol/L group;(76.0 ± 1.0) ％ in the 2.00 mmol/L group;(58.1 ± 2.6) ％ in the 4.00 mmol/L group;(52.7 ± 1.5) ％ in the 5.00 mmol/L group;(37.7 ± 3.2) ％ in the 8.00 mmol/L group;(31.3 ±2.0)％ in the 10.00 mmol/L group.The isolated exosomes demonstrated round or oval shape under TEM.The peak particle size was 56 nm,which the overall mean particle size was 87 nm.Those and particles with a diameter between 30-150 nm accounted for 91.2％.In this experiment,The expression of HSP70,CD63 could be detected in the isolated exosomes.However,only the expression of HSP70 could be detected in the HK-2 cells.Conclusions Under the treatment of 2 mmol/l oxalate for 48 hours,Ultracentrifugation can be used to isolate and purify exosomes efficiently from the HK-2 cells.This is helpful for further study of exosome as mediator of cell-to-cell communication.
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Objective To investigate the tendency of distribution and drug-resistance of the causative organisms of urinary tract infections(UTIs)in Wuhan,and provide reliable evidence for clinical treatment.Methods Analyzed the 5 378 stains of pathogen isolated from the urine of patients in hospital.The bacteria isolates were identified with BD Phoenix-100 while can-dida isolates were identified by color plate.Results A total of 5 378 stains of pathogen had been isolated.There were 2 945 stains (54.8%)of Gram-negative bacteria,1 657 stains (30.8%)of Gram-positive bacteria,776 stains (14.4%)of fungus. The rates of Escherichiacoli resistant to penicillin were highest (>83%),and there were no carbapenem-resistant strains. There were vancomycin and linezolid-resistant Enterococcispp strains,the lowest dection rates of which were 0.3%.The de-tection rate of MRCNS was over 83%.Conclusion Escherichiacoli was the most common pathogens of urinary tract infec-tion,and theβ-lactamase inhibitor complex can be used as empirical treatment of E.coli infections.Thedetection rate of MRCNS increased,which shoud be kept a watchful eye on.